Fig. 2
Characterization of HLA-A2 and HLA-A11 restricted HPV18 E7-specific CD8 + T cell responses generated by therapeutic HPV vaccine in human MHC class I transgenic mice. Female HLA-A11 or HLA-A2 transgenic mice (5 per group) were vaccinated as described in Fig. 1. For A, C 12 days after the final vaccination, the splenocytes from the mice were prepared and stimulated with HPV18 E7.1 peptide, HPV18 E7.2 peptide, HPV18 E7aa6-14 peptide, or HPV18E7aa7-15 peptide at the presence of GolgiPlug overnight. The cells were then stained with PE-conjugated anti-mouse CD8a. After permeabilization and fixation, the cells were stained with FITC-conjugated anti-mouse IFN-γ.The cells were acquired with FACSCalibur flow cytometer and data were analyzed with CellQuest Pro software. For figure B, 12 days after the last vaccination, the splenocytes from the mice were prepared and stimulated with C1R-Db, C1R-Kb, or C1R-AAD cells pulsed with HPV18 E7aa7-15 peptide. For figure D, the splenocytes were stimulated with T2-Db, T2-Kb, or T2-A11 cells pulsed with HPV18 E7aa6-14 peptide at the presence of GolgiPlug overnight. The cells were then stained as described previously. A Representative image (left panel) and summary (right panel) of flow cytometry analysis of HPV18 E7-specific CD8 + T cells generated by vaccinated HLA-A2 transgenic mice. B Representative image (left panel) and summary (right panel) of flow cytometry analysis following pulsing with C1R-Db, C1R-Kb, or C1R-AAD cells pulsed with HPV18 E7aa7-15. C Representative image (left panel) and summary (right panel) of flow cytometry analysis of HPV18 E7-specific CD8 + T cells generated by vaccinated HLA-A11 transgenic mice. D Representative image (left panel) and summary (right panel) of flow cytometry analysis following pulsing with T2-Db, T2-Kb or T2-A11 cells pulsed with HPV18 E7aa6-14